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Human Activity-Optimized CRISPR Knockout Library (3 sub-libraries in lentiCRISPRv1)
(Pooled Library #1000000100)

  • Purpose

    The Sabatini/Lander CRISPR pooled library is optimized for cleavage activity, in order to maximize the likelihood of gene knockout.

    The library is split into three sub-libraries. Two tubes each contain 90,000 gRNAs each or 5 sgRNAs per gene. The third tube contains 5,000 gRNAs.

  • Vector Backbone

    lentiCRISPR v1- Note: This plasmid has been discontinued, as an updated version is available. For confirmation of screen hits, Addgene encourages users to use the updated lentiCRISPR v2, which can be requested separately.

Ordering

Item Catalog # Description Quantity Price (USD)
Pooled Library 1000000100 Three gRNA pooled sub-libraries in lentiCRISPRv1 1 $400 Add to Cart
This material is available to academics and nonprofits only.

Library Details

  • Species
    Human
  • Genes targeted
    18,663
  • gRNAs
    187,535
  • Controls
    1000 non-targeting, 500 intergenic
  • Lentiviral Generation
    3rd

Library Shipping

This library will be delivered as three pooled DNA sub-libraries in microcentrifuge tubes on blue ice. The tubes' contents will not necessarily be frozen. For best results, minimize freeze-thaw cycles.

Note that each subpool must be transformed separately.

You will receive two sub-libraries with 90,000 gRNAs each at

  • Volume
    ∼15µL
  • Concentration
    10ng/µL

You will additionally receive one sub-library (hL3C9) with 5,000 gRNAs each at

  • Volume
    ∼20µL
  • Concentration
    20ng/µL

Depositor Comments

The depositing lab notes that the DNA preps should be performed separately and then mixed together during virus production at ratios consistent with the pool size.

Screening process for Sabatini CRISPR knockout library

Diagram of Library Screening Process

When performing sgRNA validation in a Cas9-expressing cell line, the depositing laboratory recommends using plasmid pLenti-sgRNA (Addgene #71049).

Amplified grna population graph

sgRNA diversity after initial amplification

initial grna population graph
sgRNA diversity after infection of a target cell population
How to cite this pooled library (Back to top )

These pooled libraries were created by your colleagues. Please acknowledge the Principal Investigator, cite the article in which the pooled libraries were described, and include Addgene in the Materials and Methods of your future publications.

  • Example for your Materials & Methods section:

    Activity-Optimized Human CRISPR Pooled Library was a gift from David Sabatini and Eric Lander (Addgene # 1000000100)
  • For your References section:

    Identification and characterization of essential genes in the human genome. Wang T, Birsoy K, Hughes NW, Krupczak KM, Post Y, Wei JJ, Lander ES, Sabatini DM. Science. 2015 Oct 15. pii: aac7041. PubMed 26472758